<2022-01-04 Tue> David biweekly meeting

projects

Project 4: Reconstitution of endocytic actin network

recent progress

  • successfully correlated actin-positive beads between fluorescence and EM

    • light microscope field (bright field, Sac6-GFP, far-red lipid)

      2021-12-07_11-34-30_Screen Shot 2021-12-06 at 1.24.48 PM.png

    • bead + actin at top right:

      2021-12-07_11-37-51_Screen Shot 2021-12-06 at 1.27.08 PM.png

    • bead with very low actin signal at middle left:

      2021-12-07_11-38-44_Screen Shot 2021-12-06 at 1.37.22 PM.png
  • some interesting membrane structures on Sac6-GFP enriched beads

    • vesicle in actin

      2022-01-04_13-13-25_Screen Shot 2022-01-04 at 1.09.42 PM.png
      2022-01-04_13-13-37_Screen Shot 2022-01-04 at 1.10.01 PM.png

    • big old vesicle inside actin, lots of bumps on bead surface

      2022-01-04_13-14-10_Screen Shot 2022-01-04 at 1.10.43 PM.png

    • some membrane structures seem to be blasting off

      2022-01-04_13-15-03_Screen Shot 2022-01-04 at 1.11.07 PM.png

    • a bigger vesicle with maybe an actin tail

      2022-01-04_13-16-32_Screen Shot 2022-01-04 at 1.11.58 PM.png
      2022-01-04_13-16-46_Screen Shot 2022-01-04 at 1.12.21 PM.png

    • another structure blasting off

      2022-01-04_13-17-15_Screen Shot 2022-01-04 at 1.12.37 PM.png
  • currently communicating with tech support to fix Wall-E laser, not sure how long this will take
  • in the meantime, I ordered a different lipid fluorescent label and will get trained on a different MIC confocal scope that should have the features I need

next steps

  • fix wall-E and image negative control beads for correlation
  • determine whether there is a difference in membrane shape between high and low fluorescence beads
    • come up with some metric of scoring if so
  • if there are differences, collect triplicate data and do statistical test

Project 7: Mechanical regulation of CME by protein LLPS

recent progress

  • figured out how to run simulations on lab workstation
  • the membrane-only simulation is fully working now

    • with only membrane tension and bending energy, it relaxes to flat

    • with turgor pressure added (no cell wall), it bows outward

  • starting to set up a protein (e.g. clathrin)-coated region:

next steps

short-term
  • add cell wall to model
  • correctly add clathrin coat to model
  • add droplet to model

Date: \today

Author: Max Ferrin

Email: ferrinm@berkeley.edu

Created: 2022-01-04 Tue 14:24

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